| Objective:Parkinson's disease(PD)is a disease with abnormal neuron apoptosis as the main pathogenesis.Studies at home and abroad have shown that moxibustion can effectively prevent and delay the process of PD.The mechanism may be that moxibustion can protect neurons from neuronal apoptosis.The PI3K/Akt signaling pathway is an important pathway to promote cell survival.Is the mechanism of moxibustion prevention and treatment of PD related to the PI3K/Akt signaling pathway? In this experiment,the insecticide rotenone was used as a model inducer.The test object was rotenone-induced PD model rats.The guiding ideology is the theory of traditional Chinese medicine and moxibustion was used as the treatment method.Select "Guan Yuan(CV4),Zusanli(ST36),Hefengfu(GV16)" three points;taking anti-neuronal apoptosis as the entry point;observing the effect of moxibustion on the number of TH and TH neurons and the expression of PI3K and Akt in the substantia nigra of PD model rats,and exploring moxibustion treatment The possible mechanism of PD is to provide more experimental evidence for the wider clinical use of moxibustion in the prevention and treatment of PD.Methods: Fifty 6-week-old SPF-grade healthy male SD rats weighing 180-220 g had undergone three behavioral tests and had no spontaneous behavioral abnormalities.The above rats were included in the experimental study.After 14 days of adaptive feeding in each group of rats,then 20 rats were drawn and divided into two groups of 10 rats each.The rats were grouped according to the random number table method,that is the normal group and the sham operation group,the remaining rats(30 rats)were subcutaneously injected with rotenone solution(rotenone was dissolved in a mixed solution of dimethyl sulfoxide and normal saline,in which the volume ratio of dimethyl sulfoxide to normal saline was 7: 1,and the concentration of rotenone was 1mg/ml,rat injection dose was 1ml/kg).The sham operation group was injected with a mixed solution of dimethanfeng and normal saline at the same site(volume ratio 7: 1,injection volume was 1ml/kg).The normal group was not given any treatment,and only grasped and fixed once a day;the above operation was performed once a day for a total of 40 days.At 9 am on the second day after the end of modeling,the appearance and behavioral performance of each group of rats were scored with reference to the behavioral scoring standards established by Chen Xin and others.Rats with a score of 2-8 were regarded as successful in modeling and excluded.Unsuccessful rat modeling.The 20 rats successfully modeled were randomly divided into a model group and a treatment group.The rats in the treatment group were fixed with simple clothing.With reference to the method of acupoints in Experimental Acupuncture,select "Guan Yuan(CV4),Zu Sanli(ST36)and Fengfu(GV16)" three acupuncture points,using moxa sticks with a length of 10 cm and a diameter of 0.7cm are ignited about 2cm above the acupuncture point(the specific height is adjusted slightly based on the temperature of the local skin touched by the moxibustion and the performance of the rat during the moxibustion.),Moxibustion at each acupoint for 15min(the use of Zusanli on both sides(ST36)alternately),once a day,continuous treatment for 14 days;no treatment was given to the remaining 3 groups of rats during the treatment period,only grasp and fix at the same time every day.Rats of each group were fed synchronously during the entire modeling and treatment period.After that,the behavioral scores of each group of rats were statistically compared.After the behavioral scores were completed,experimental samples were obtained from the rats in each group(midbrain nigra): half of the rats in each group underwent left ventricular paraformaldehyde perfusion and internal fixation,and brain tissue was taken as sections to detect the positive expression of TH and TH neurons in substantia nigra of rats.After the rats in each of the remaining groups were anesthetized by intraperitoneal injection,fresh brain tissue was homogenized.The relative expression levels of PI3K mRNA and Akt mRNA and the protein expression in the substantia nigra of each group were detected for statistical analysis and comparison.Results:(1)Compared with the normal group and the sham operation group,the appearance and behavior scores of the model group were significantly increased(P<0.01);after 14 days of moxibustion treatment,compared with the model group,the appearance and behavior of the treatment group rats The academic performance was significantly improved,and the score was significantly decreased(P<0.01).(2)Compared with the normal group and the sham operation group,the TH immune response in the substantia nigra of the model group was weakened,and the average optical density value was significantly reduced(P<0.05).After 14 days of moxibustion treatment,compared with the model group,the treatment The TH immune response and mean optical density of the rats in the group were significantly increased(P<0.05).(3)Compared with the normal group and the sham operation group,the number of substantia nigra TH neurons in the model group was significantly decreased(P<0.01).After 14 days of moxibustion treatment,compared with the model group,the substantia nigra TH neurons in the treatment group increased significantly(P<0.01).(4)Compared with normal group and sham operation group,the expression of substantia nigra PI3K and Akt protein in model group rats was decreased(P<0.05).After 14 days of moxibustion treatment,compared with model group,substantia nigra PI3K and Akt protein expression increased significantly(P<0.05).(5)Compared with the normal group and the sham operation group,the relative expression of PI3K mRNA and Akt mRNA in the substantia nigra of the model group was decreased(P<0.05).After 14 days of moxibustion treatment,compared with the model group,The relative expression of PI3K mRNA and Akt mRNA increased significantly(P<0.05).Conclusions:(1)Subcutaneous injection of rotenone on the back of the neck can successfully replicate the PD model.(2)The appearance and behavior abnormalities of model rats can be significantly improved by moxibustion treatment.(3)The content of TH in the substantia nigra of the model group rats was reduced,and it was significantly increased after moxibustion treatment,and the number of TH neurons was increased.(4)Moxibustion treatment can significantly increase the relative expression of PI3K mRNA and Akt mRNA and their protein expression levels in substantia nigra of PD model rats,indicating that moxibustion may activate the PI3K/Akt signaling pathway,thereby exerting inhibition of neuronal apoptosis and improving PD effects of symptoms in Model Rats. |
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